Single-step, wash-free digital immunoassay for rapid quantitative analysis of serological antibody against SARS-CoV-2 by photonic resonator absorption microscopy.
Single-step, wash-free digital immunoassay for rapid quantitative analysis of serological antibody against SARS-CoV-2 by photonic resonator absorption microscopy.
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DOI：
10.1016/j.talanta.2020.122004
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发表时间：
2021-04-01
期刊：
影响因子：
6.1
通讯作者：
Cunningham BT
中科院分区：
文献类型：
--
作者：
Zhao B;Che C;Wang W;Li N;Cunningham BT
严重急性呼吸系统综合征冠状病毒2型（SARS-CoV-2）是2019年冠状病毒病（COVID-19）的病因，对全球公共卫生构成了极大的威胁和复杂的挑战。SARS-CoV-2抗体滴度的定量测定在了解患者间免疫应答的变异性、评估疫苗的有效性以及识别输血治疗的供体方面起着重要作用。对高度灵敏、定量、快速、简单、微创和廉价的血清学COVID-19抗体检测的需求迫切且不断增长。在这项工作中，我们开发了一个单步，免洗涤免疫分析快速和高灵敏度的定量分析血清学人IgG抗SARS-CoV-2，只需要一个单一的血清液滴。通过简单地将4 μL人血清样品与抗体功能化的金纳米颗粒孵育，涂覆有重组spike蛋白的光子晶体光学生物传感器作为传感平台，用于通过特异性抗原-抗体相互作用形成夹心免疫复合物，在该传感平台上，检测到的IgG分子可以以数字精度计数。我们证明了一种单步15分钟测定法，能够检测血清样本中低至100 pg mL−1的人COVID-19 IgG。计算的检测限（LOD）和定量限（LOQ）分别为26.7 ± 7.7和32.0 ± 8.9 pg mL−1。这项工作代表了首次利用基于Activate Capture + Digital Counting（AC + DC）的免疫测定法对血清学COVID-19抗体进行快速定量分析，展示了使用便携式检测仪器进行即时检测的途径。基于夹心免疫测定原理，生物传感平台可以扩展用于抗原、额外IgG、细胞因子和其他蛋白质生物标志物的多重检测。基于PRAM的血清学COVID-19 IgG数字免疫分析。
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the cause of Coronavirus Disease 2019 (COVID-19), poses extraordinary threats and complex challenges to global public health. Quantitative measurement of SARS-CoV-2 antibody titer plays an important role in understanding the patient-to-patient variability of immune response, assessing the efficacy of vaccines, and identifying donors for blood transfusion therapy. There is an urgent and ever-increasing demand for serological COVID-19 antibody tests that are highly sensitive, quantitative, rapid, simple, minimally invasive, and inexpensive. In this work, we developed a single-step, wash-free immunoassay for rapid and highly sensitive quantitative analysis of serological human IgG against SARS-CoV-2 which requires only a single droplet of serum. By simply incubating 4 μL human serum samples with antibody-functionalized gold nanoparticles, a photonic crystal optical biosensor coated with the recombinant spike protein serves as a sensing platform for the formation of sandwich immunocomplex through specific antigen–antibody interactions, upon which the detected IgG molecules can be counted with digital precision. We demonstrated a single-step 15-min assay capable of detecting as low as 100 pg mL−1 human COVID-19 IgG in serum samples. The calculated limit of detecting (LOD) and limit of quantification (LOQ) is 26.7 ± 7.7 and 32.0 ± 8.9 pg mL−1, respectively. This work represents the first utilization of the Activate Capture + Digital Counting (AC + DC)-based immunoassay for rapid and quantitative analysis of serological COVID-19 antibody, demonstrating a route toward point-of-care testing, using a portable detection instrument. On the basis of the sandwich immunoassay principle, the biosensing platform can be extended for the multiplexed detection of antigens, additional IgGs, cytokines, and other protein biomarkers. PRAM-based Digital Immunoassay for Serological COVID-19 IgG.
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DOI：
10.1073/pnas.1904770116
发表时间：
2019-09-24
影响因子：
11.1
作者：
Canady, Taylor D.;Li, Nantao;Cunningham, Brian T.
通讯作者：
Cunningham, Brian T.